Monospecific and multispecific antibodies and method of use
View Patent ↗This invention relates to monospecific and multispecific antibodies that may be utilized for the diagnosis and treatment of various diseases. In addition, these antibodies may be modified by protease cleavage. Protease control or regulation may be provided by a protease site located in, for example, a linker. These protease-regulated antibodies may also be utilized for the diagnosis and treatment of various diseases.
1. An antibody comprising a heavy chain and a light chain, each comprising one or more variable regions, wherein said antibody is capable of binding an antigen or an epitope, and wherein said antibody comprises an amino acid linker which comprises one or more protease cleavage sites, wherein said protease cleavage sites are selected from the group consisting of SEQ ID NOS:26, 27, 28, 29, 30, 31, and 32, wherein the antibody comprises an amino acid sequence selected from the group consisting of SEQ ID NO:33-120.
2. A kit comprising an antibody which comprises a heavy chain and a light chain, each comprising one or more variable regions, wherein the antibody is capable of binding an antigen or an epitope, and wherein the antibody comprises an amino acid linker which comprises one or more protease cleavage sites, wherein the protease cleavage sites are selected from the group consisting of SEQ ID NOS:26, 27, 28, 29, 30, 31, and 32, wherein the antibody comprises (a) a heavy chain having the structure NH 2 -V H 1-linker-V H 2-C H 1-CH2-CH3-COOH and (b) a light chain having the structure NH 2 -V L 1-linker-V L 2-CL-COOH, and wherein the light chain comprises the amino acid sequence SEQ ID NO:81.
3. A kit comprising an antibody which comprises a heavy chain and a light chain, each comprising one or more variable regions, wherein the antibody is capable of binding an antigen or an epitope, and wherein the antibody comprises an amino acid linker which comprises one or more protease cleavage sites, wherein the protease cleavage sites are selected from the group consisting of SEQ ID NOS:26, 27, 28, 29, 30, 31, and 32, wherein the antibody comprises (a) a heavy chain having the structure NH 2 -V H 1-linker-V H 2-C H 1-CH2-CH3-COOH and (b) a light chain having the structure NH 2 -V L 1-linker-V L 2-CL-COOH, and wherein the heavy chain comprises the amino acid sequence SEQ ID NO:82.
4. A kit comprising an antibody which comprises a heavy chain and a light chain, each comprising one or more variable regions, wherein the antibody is capable of binding an antigen or an epitope, and wherein the antibody comprises an amino acid linker which comprises one or more protease cleavage sites, wherein the protease cleavage sites are selected from the group consisting of SEQ ID NOS:26, 27, 28, 29, 30, 31, and 32, wherein the antibody comprises (a) a heavy chain having the structure NH 2 -V H 1-linker-V H 2-C H 1-CH2-CH3-COOH and (b) a light chain having the structure NH 2 -V L 1-linker-V L 2-CL-COOH, and wherein the light chain comprises the amino acid sequence SEQ ID NO:81 and the heavy chain comprises the amino acid sequence SEQ ID NO:82.
5. An antibody which comprises a heavy chain and a light chain, each comprising one or more variable regions, wherein the antibody is capable of binding an antigen or an epitope, and wherein the antibody comprises an amino acid linker which comprises one or more protease cleavage sites, wherein the protease cleavage sites are selected from the group consisting of SEQ ID NOS:26, 27, 28, 29, 30, 31, and 32, wherein the antibody comprises (a) a heavy chain having the structure NH 2 -V H 1-linker-V H 2-C H 1-CH2-CH3-COOH and (b) a light chain having the structure NH 2 -V L 1-linker-V L 2-CL-COOH, and wherein the light chain comprises the amino acid sequence SEQ ID NO:81.
6. An antibody which comprises a heavy chain and a light chain, each comprising one or more variable regions, wherein the antibody is capable of binding an antigen or an epitope, and wherein the antibody comprises an amino acid linker which comprises one or more protease cleavage sites, wherein the protease cleavage sites are selected from the group consisting of SEQ ID NOS:26, 27, 28, 29, 30, 31, and 32, wherein the antibody comprises (a) a heavy chain having the structure NH 2 -V H 1-linker-V H 2-C H 1-CH2-CH3-COOH and (b) a light chain having the structure NH 2 -V L 1-linker-V L 2-CL-COOH, and, wherein the heavy chain comprises the amino acid sequence SEQ ID NO:82.
7. An antibody which comprises a heavy chain and a light chain, each comprising one or more variable regions, wherein the antibody is capable of binding an antigen or an epitope, and wherein the antibody comprises an amino acid linker which comprises one or more protease cleavage sites, wherein the protease cleavage sites are selected from the group consisting of SEQ ID NOS:26, 27, 28, 29, 30, 31, and 32, wherein the antibody comprises (a) a heavy chain having the structure NH 2 -V H 1-linker-V H 2-C H 1-CH2-CH3-COOH and (b) a light chain having the structure NH 2 -V L 1-linker-V L 2-CL-COOH, and, wherein the light chain comprises the amino acid sequence SEQ ID NO:81 and the heavy chain comprises the amino acid sequence SEQ ID NO:82.
8. The antibody of claim 5 which is conjugated to monomethylauristatin-E (MMAE).
9. The antibody of claim 6 which is conjugated to monomethylauristatin-E (MMAE).
10. The antibody of claim 7 which is conjugated to monomethylauristatin-E (MMAE).
11. A pharmaceutical composition comprising the antibody of claim 5 .
12. A pharmaceutical composition comprising the antibody of claim 6 .
13. A pharmaceutical composition comprising the antibody of claim 7 .
14. The pharmaceutical composition of claim 11 , wherein the antibody is conjugated to monomethylauristatin-E (MMAE).
15. The pharmaceutical composition of claim 12 , wherein the antibody is conjugated to monomethylauristatin-E (MMAE).
16. The pharmaceutical composition of claim 13 , wherein the antibody is conjugated to monomethylauristatin-E (MMAE).
17. A kit comprising:
an antibody comprising one or more variable regions, wherein each of the one or more variable regions comprises a heavy chain and a light chain and is capable of binding an antigen or an epitope, and wherein the antibody comprises an amino acid linker which comprises one or more protease cleavage sites; and
solutions for suspending or fixing the cells, detectable labels, solutions for rendering a polypeptide susceptible to the binding of an antibody, solutions for lysing cells, and/or solutions for the purification of polypeptides
wherein the antibody comprises a heavy chain having the structure NH 2 -V H 1-linker-V H 2-C H 1-CH2-CH3-COOH; and a light chain having the structure NH 2 -V L 1-linker-V L 2-CL-COOH, and wherein the light chain comprises the amino acid sequence SEQ ID NO:81.
18. A kit comprising:
an antibody comprising one or more variable regions, wherein said antibody binds one or more antigens or epitopes; and
solutions for suspending or fixing the cells, detectable labels, solutions for rendering a polypeptide susceptible to the binding of an antibody, solutions for lysing cells, and/or solutions for the purification of polypeptides
wherein the antibody comprises a heavy chain having the structure NH 2 -V H 1-linker-V H 2-C H 1-CH2-CH3-COOH; and a light chain having the structure NH 2 -V L 1-linker-V L 2-CL-COOH, and wherein the heavy chain comprises the amino acid sequence SEQ ID NO:82.
19. The kit of claim 18 , wherein the light chain comprises the amino acid sequence SEQ ID NO:81.
20. An antibody comprising one or more variable regions, wherein said antibody binds one or more antigens or epitopes, wherein said variable region comprises a heavy chain having the structure NH 2 -V H 1-linker-V H 2-C H 1-CH2-CH3-COOH; and a light chain having the structure NH 2 -V L 1-linker-V L 2-CL-COOH, and wherein said antibody comprises a variable region light chain comprising the amino acid sequence SEQ ID NO:81.
21. An antibody comprising one or more variable regions, wherein said antibody binds one or more antigens or epitopes, wherein said variable region comprises a heavy chain having the structure NH 2 -V H 1-linker-V H 2-C H 1-CH2-CH3-COOH; and a light chain having the structure NH 2 -V L 1-linker-V L 2-CL-COOH, and wherein said antibody comprises a variable region heavy chain comprising the amino acid sequence SEQ ID NO:82.
22. The antibody of claim 21 , wherein said antibody comprises a variable region light chain comprising the amino acid sequence SEQ ID NO:81.
23. The antibody of claim 20 , which is conjugated to monomethylauristatin-E (MMAE).
24. The antibody of claim 21 , which is conjugated to monomethylauristatin-E (MMAE).
25. The antibody of claim 22 , which is conjugated to monomethylauristatin-E (MMAE).
26. A pharmaceutical composition, comprising an antibody in combination with a pharmaceutically acceptable carrier, wherein the antibody comprises one or more variable regions, wherein said antibody binds one or more antigens or epitopes, wherein said variable region comprises a heavy chain having the structure NH 2 -V H 1-linker-V H 2-C H 1-CH2-CH3-COOH; and a light chain having the structure NH 2 -V L 1-linker-V L 2-CL-COOH, and wherein said antibody comprises a variable region light chain comprising the amino acid sequence SEQ ID NO:81.
27. A pharmaceutical composition, comprising an antibody in combination with a pharmaceutically acceptable carrier, wherein the antibody comprises one or more variable regions, wherein said antibody binds one or more antigens or epitopes, wherein said variable region comprises a heavy chain having the structure NH 2 -V H 1-linker-V H 2-C H 1-CH2-CH3-COOH; and a light chain having the structure NH 2 -V L 1-linker-V L 2-CL-COOH, and wherein said antibody comprises a variable region heavy chain comprising the amino acid sequence SEQ ID NO:82.
28. The pharmaceutical composition of claim 27 , wherein the antibody comprises a variable region light chain comprising the amino acid sequence SEQ ID NO:81.
29. The pharmaceutical composition of claim 26 , wherein the antibody is conjugated to monomethylauristatin-E (MMAE).
30. The pharmaceutical composition of claim 27 , wherein the antibody is conjugated to monomethylauristatin-E (MMAE).
31. The pharmaceutical composition of claim 28 , wherein the antibody is conjugated to monomethylauristatin-E (MMAE).
32. The antibody of claim 1 , wherein said antibody is conjugated to a therapeutic or cytotoxic agent.
33. The antibody of claim 32 , wherein said antibody is conjugated to an agent selected from the group consisting of monomethylauristatin-E (MMAE), aplidin, azaribine, anastrozole, azacytidine, bleomycin, bortezomib, bryostatin-1, busulfan, calicheamycin, camptothecin, 10-hydroxycamptothecin, carmustine, celebrex, chlorambucil, cisplatin, irinotecan (CPT-I1), SN-38, carboplatin, cladribine, cyclophosphamide, cytarabine, dacarbazine, docetaxel, dactinomycin, daunomycin glucuronide, daunorubicin, dexamethasone, diethylstilbestrol, doxorubicin, doxorubicin glucuronide, epirubicin glucuronide, ethinyl estradiol, estramustine, etoposide, etoposide glucuronide, etoposide phosphate, floxuridine (FUdR), 3′,5′-O-dioleoyl-FudR (FUdR-dO), fludarabine, flutamide, fluorouracil, fluoxymesterone, gemcitabine, hydroxyprogesterone caproate, hydroxyurea, idarubicin, ifosfamide, L-asparaginase, leucovorin, lomustine, mechlorethamine, medroprogesterone acetate, megestrol acetate, melphalan, mercaptopurine, 6-mercaptopurine, methotrexate, mitoxantrone, mithramycin, mitomycin, mitotane, phenyl butyrate, prednisone, procarbazine, paclitaxel, pentostatin, PSI-341, semustine streptozocin, tamoxifen, taxanes, taxol, testosterone propionate, thalidomide, thioguanine, thiotepa, teniposide, topotecan, uracil mustard, velcade, vinblastine, vinorelbine, vincristine, ricin, abrin, ribomiclease, onconase, rapLR1, DNase I, Staphylococcal enterotoxin-A, pokeweed antiviral protein, gelonin, diphtheria toxin, Pseudomonas exotoxin, and Pseudomonas endotoxin, and functional analogs thereof.
34. A pharmaceutical composition comprising a therapeutically effective amount of an antibody of claim 1 in combination with a pharmaceutically acceptable carrier.
35. The pharmaceutical composition of claim 34 further comprising one or more pharmaceutical agents.
36. A kit comprising an antibody of claim 1 .
37. The kit of claim 36 , further comprising solutions for suspending or fixing the cells, detectable labels, solutions for rendering a polypeptide susceptible to the binding of an antibody, solutions for lysing cells, and/or solutions for the purification of polypeptides.
38. The kit of claim 37 wherein the antibody comprises a heavy chain having the structure NH 2 -V H 1-linker-V H 2-C H 1-CH2-CH3-COOH; and a light chain having the structure NH 2 -V L 1-linker-V L 2-CL-COOH.